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  • C07D307/91—Dibenzofurans; Hydrogenated dibenzofurans
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  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/04—Antibacterial agents
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  • C07C57/46—Unsaturated compounds having carboxyl groups bound to acyclic carbon atoms containing six-membered aromatic rings and other rings, e.g. cyclohexylphenylacetic acid
  • C07C57/48—Unsaturated compounds having carboxyl groups bound to acyclic carbon atoms containing six-membered aromatic rings and other rings, e.g. cyclohexylphenylacetic acid having unsaturation outside the aromatic rings
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  • C07C57/52—Unsaturated compounds having carboxyl groups bound to acyclic carbon atoms containing halogen
  • C07C57/58—Unsaturated compounds having carboxyl groups bound to acyclic carbon atoms containing halogen containing six-membered aromatic rings
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  • C07C59/40—Unsaturated compounds
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  • C07C59/40—Unsaturated compounds
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  • C07C59/40—Unsaturated compounds
  • C07C59/58—Unsaturated compounds containing ether groups, groups, groups, or groups
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  • C07C59/40—Unsaturated compounds
  • C07C59/58—Unsaturated compounds containing ether groups, groups, groups, or groups
  • C07C59/64—Unsaturated compounds containing ether groups, groups, groups, or groups containing six-membered aromatic rings
  • C07C59/66—Unsaturated compounds containing ether groups, groups, groups, or groups containing six-membered aromatic rings the non-carboxylic part of the ether containing six-membered aromatic rings
  • C07C59/68—Unsaturated compounds containing ether groups, groups, groups, or groups containing six-membered aromatic rings the non-carboxylic part of the ether containing six-membered aromatic rings the oxygen atom of the ether group being bound to a non-condensed six-membered aromatic ring
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  • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
  • C07D209/56—Ring systems containing three or more rings
  • C07D209/80—[b, c]- or [b, d]-condensed
  • C07D209/82—Carbazoles; Hydrogenated carbazoles
  • C07D209/86—Carbazoles; Hydrogenated carbazoles with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to carbon atoms of the ring system
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  • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
  • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
  • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
  • C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
  • C07D213/54—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
  • C07D213/55—Acids; Esters
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  • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
  • C07D307/02—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings
  • C07D307/34—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
  • C07D307/38—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with substituted hydrocarbon radicals attached to ring carbon atoms
  • C07D307/54—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
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  • C07D317/00—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms
  • C07D317/08—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3
  • C07D317/44—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3 ortho- or peri-condensed with carbocyclic rings or ring systems
  • C07D317/46—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3 ortho- or peri-condensed with carbocyclic rings or ring systems condensed with one six-membered ring
  • C07D317/48—Methylenedioxybenzenes or hydrogenated methylenedioxybenzenes, unsubstituted on the hetero ring
  • C07D317/50—Methylenedioxybenzenes or hydrogenated methylenedioxybenzenes, unsubstituted on the hetero ring with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to atoms of the carbocyclic ring
  • C07D317/60—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
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  • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
  • C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
  • C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
  • C07D333/06—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to the ring carbon atoms
  • C07D333/24—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
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  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
  • C07D333/50—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom condensed with carbocyclic rings or ring systems
  • C07D333/76—Dibenzothiophenes

Definitions

• the present invention relates to compounds which have metallo- ⁇ - lactamase inhibitory characteristics.
• the invention also relates to methods of preparing, pharmaceutical compositions and uses of the compounds.
• Metallo- ⁇ -lactamases are bacterial enzymes which confer resistance to virtually all clinically relevant ⁇ -lactam antibiotics, including carbapenems and jeopardize the future use of all such agents.
• the increased treatment of infections with carbapenems and other ⁇ -lactam antibiotics may lead to the proliferation of clinical bacterial strains which are able to produce metallo- ⁇ -lactamases and thus resist the effects of ⁇ -lactam antibiotics.
• metallo- ⁇ -lactamases have now been identified in a number of pathogenic bacterial species including Bacillus cereus,
• WO 98/17639, 97/30027, 98/40056,98/39311 and 97/10225 teach certain beta-thiopropionyl-amino acid derivatives and their use as inhibitory agents against metallo- ⁇ -lactamases. Goto et. al, Biol. Pharm. Bull. 20, 1136 (1997), Payne et. al., FEMS Microbiology Letters 157, 171 (1997), Payne et al., Antimicrob. Agents Chemother. 41, 135 (1997), Page et. al, Chem. Commun. 1609 (1998) and Page et al., Biochem. J. 331, 703 (1998) also disclose certain thiols and thioesters as metallo- ⁇ - lactamase inhibitors. Additionally, Toney et al., Chemistry and Biology 5, 185
• This invention relates to novel substituted succinic acid metallo- ⁇ - lactamase inhibitors, which are useful potentiators of ⁇ -lactam antibiotics.
• the present invention provides a method of treating bacterial infections in animals or humans which comprises administering, together with a ⁇ -lactam antibiotic, a therapeutically effective amount of a compound of formula I:
• Ml and M-2 are independently selected from:
• R 1 and R 2 are independently selected from the following:
• -A- represents a single bond, Cl to C$ straight, branched or unsaturated alkyl group optionally substituted with 1 to 2 R x groups and optionally interrupted by one of the following O, S, SO2, -C(O)-, -C(O)-NRa-, -CO 2 -;
• A' is a single bond, O, S, or a Cl to C6 straight, branched or unsaturated alkyl group optionally substituted with 1 to 2 R x groups and optionally interrupted by one of the following groups O, S, SO2, -C(O)-, -C(O)-NRa_, -CO 2 -;
• B J and C ⁇ are independently selected from: (1) a C 6 to C-o aryl group;
• each R x is independently selected from the group consisting of: (a) F, Cl, Br, I , (b) CF 3 ,
• Ra is H, Cl to C6 alkyl optionally substituted with RY;
• Rb is H, Cl to C6 alkyl optionally substituted with Ry, CH2-aryl, or aryl, said aryls optionally substituted with 1-2 RY groups;
• R c is H, Cl to C6 alkyl optionally substituted with RY, CF3, or aryl, said aryl optionally substituted with 1 to 2 RY groups;
• Rd and R e are independently hydrogen, Cl to C4 alkyl optionally substituted with RY. or Rd and R e taken together may represent a 3 to 5-membered alkyl radical to form a ring, or Rd and Re taken together may represent a 2 to 4-membered alkyl radical interrupted by O, S, SO or SO2 to form a ring;
• Rf is Cl to C6 alkyl optionally substituted with RY, or aryl, said aryl optionally substituted with 1 to 2 RY groups;
• RY is -OH, -OCH3, OCONH2, OCOCH3, CHO, COCH3, CO2CH3, CONH2, CN, SOCH3, SO2CH3, SO2NH2, F, Cl, Br, I or CF3.
• the invention is intended to include all of the isomeric forms of the compounds of formula I, including racemic, enantiomeric and diastereomeric forms.
• compositions containing the compounds of formula I and method of treatments using the same are also included in this invention.
• alkyl refers to a monovalent alkane (hydrocarbon) derived radical containing from 1 to 16 carbon atoms unless otherwise defined. It may be straight or branched. Preferred alkyl groups include methyl, ethyl, propyl, isopropyl, butyl and hexyl. When substituted, alkyl groups may be substituted with up to 3 substituent groups, selected from Rx as defined, at any available point of attachment. When the alkyl group is said to be substituted with an alkyl group, this is used interchangeably with “branched alkyl group”. When the alkyl chain is interrupted by a group, eg. O, this may occur between any two saturated carbons of the alkyl chain.
• unsaturated alkyl refers to "alkenyl” or "alkynyl”.
• alkenyl refers to an unsaturated alkyl such as a hydrocarbon radical, straight or branched containing from 2 to 16 carbon atoms and at least one carbon to carbon double bond.
• Preferred alkenyl groups include propenyl, hex enyl and butenyl.
• alkynyl refers to an unsaturated alkyl such as a hydrocarbon radical straight or branched, containing from 2 to 16 carbon atoms and at least one carbon to carbon triple bond.
• Preferred alkynyl groups include propynyl, hexynyl and butynyl.
• alicyclic refers to non-aromatic monocyclic or bicyclic C3-C10 hydrocarbons, including unsaturated, which can be substituted with 0-3 groups of R x .
• groups include cycloalkyls such as cyclohexyl, cyclopentyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]hepta-2,5-dienyl, bicyclo[2.2.2]octyl, bicyclo[2.2.2]octa-2,5-dienyl.
• alkylidene refers to an alkyl group which is attached through two bonds on the same carbon atom of the alkyl group to a single attachment atom.
• alkylidene refers to an alkyl group which is attached through two bonds on the same carbon atom of the alkyl group to a single attachment atom. Examples of said groups include methylene, ethylidene, isopropylidene and the like.
• Rd and Re are taken together along with the adjacent nitrogen atom to represent a 3 to 5 membered alkyl radical forming a ring or a 2 to 4 membered alkyl radical interrupted by O, S, SO, SO2, to form a ring are pyrrolidinyl, piperidinyl, morpholinyl and the like.
• heterocyclic refers to a monocyclic non-aromatic moiety containing 3-8 ring atoms or a bicyclic non-aromatic moiety containing 6-10 ring atoms, at least one of which ring atoms is a heteroatom selected from nitrogen, oxygen and sulfur and where one additional ring atom may be oxygen or sulfur.
• heterocyclic groups are furanyl, pyranyl, morpholinyl, dioxanyl and quinuclidinyl: furan pyran ioxane quinuclidine
• Aryl refers to aromatic rings e.g., phenyl, substituted phenyl and the like, as well as rings which are fused, e.g., naphthyl, phenanthrenyl fluorenonyl and the like.
• An aryl group thus contains at least one ring having at least 6 atoms, with up to three such rings being present, containing up to 14 atoms therein, with alternating (resonating) double bonds between adjacent carbon atoms.
• the preferred aryl groups are phenyl, naphthyl, and fluorenone.
• Aryl groups may likewise be substituted as defined.
• Preferred substituted aryls include phenyl, fluorenonyl and naphthyl.
• heteroaryl refers to a monocyclic aromatic group having 5 or 6 ring atoms, a bicyclic aromatic group having 8 to 10 atoms, or tricyclic having 12-14 ring atoms, containing at least one heteroatom, O, S or N, in which a carbon or nitrogen atom is the point of attachment, and in which one or two additional carbon atoms is optionally replaced by a heteroatom selected from O or S, and in which from 1 to 3 additional carbon atoms are optionally replaced by nitrogen heteroatoms, said heteroaryl group being optionally substituted as described herein.
• Examples of this type are pyrrole, pyridine, oxazole, thiazole, dibenzofuran, dibenzothiophene, carbazole, phenanthrene, anthracene, dibenzothiophene sulfone, fluorenone, quinoline and oxazine. Additional nitrogen atoms may be present together with the first nitrogen and oxygen or sulfur, giving, e.g., thiadiazole. Examples include the following:
• heteroatom means O, S or N, selected on an independent basis.
• Halogen and "halo” refer to bromine, chlorine, fluorine and iodine.
• pro-drug refers to compounds with a removable group attached to one or both of the carboxyl groups of compounds of formula I (e.g. biolabile esters). Groups which are useful in forming pro-drugs should be apparent to the medicinal chemist from the teachings herein. Examples include pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl and methoxymethyl, and others described in detail in U.S. Pat. No. 4,479,947. These are also referred to as "biolabile esters.
• hydrate is used in the conventional sense to include the compounds of formula I in physical association with water.
• substituted unless otherwise indicated, this means that the group contains from 1 to 3 substituents thereon.
• a bond terminated by a wavy line is used herein to signify the point of attachment of a substituent group. This usage is illustrated by the following example:
• Suitable protecting groups for the compounds of the present invention will be recognized from the present application taking into account the level of skill in the art, and with reference to standard textbooks, such as Greene, T. W. et al. Protective
• suitable protecting groups represents hydroxyl-protecting or carboxyl-protecting groups.
• Such conventional protecting groups consist of groups, which are used to protectively block the hydroxyl or carboxyl group during the synthesis procedures described herein.
• These conventional blocking groups are readily removable, i.e., they can be removed, if desired, by procedures which will not cause cleavage or other disruption of the remaining portions of the molecule.
• Such procedures include chemical and enzymatic hydrolysis, treatment with chemical reducing or oxidizing agents under mild conditions, treatment with a transition metal catalyst and a nucleophile and catalytic hydrogenation.
• carboxyl protecting groups include allyl, benzhydryl, 2-naphthylmethyl, benzyl, silyl such as t-butyldimethylsilyl (TBDMS), phenacyl, p-methoxybenzyl, o-nitrobenzyl, p-methoxyphenyl, p-nitrobenzyl, 4-pyridylmethyl and t-butyl.
• Suitable hydroxyl protecting groups include triethylsilyl, t-butyldimethylsilyl, o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, benzyloxycarbonyl, allyloxycarbonyl, t-butyloxycarbonyl, 2,2,2- trichloroethyloxycarbonyl and the like.
• the compounds of the present invention are useful per se and in their pharmaceutically acceptable salt and ester forms are potentiators for the treatment of bacterial infections in animal and human subjects.
• pharmaceutically acceptable ester, salt or hydrate refers to those salts, esters and hydrated forms of the compounds of the present invention which would be apparent to the pharmaceutical chemist, i.e., those which are substantially non-toxic and which may favorably affect the pharmacokinetic properties of said compounds, such as palatability, absorption, distribution, metabolism and excretion.
• Other factors, more practical in nature, which are also important in the selection, are cost of the raw materials, ease of crystallization, yield, stability, solubility, hygroscopicity and flowability of the resulting bulk drug.
• compositions may be prepared from the active ingredients in combination with pharmaceutically acceptable carriers.
• the present invention is also concerned with pharmaceutical compositions and methods of treating bacterial infections utilizing as an active ingredient the novel metallo-beta-lactamase inhibitors of formula I.
• the pharmaceutically acceptable salts referred to above also include acid addition salts.
• the Formula I compounds can be used in the form of salts derived from inorganic or organic acids. Included among such salts are the following: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2- hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamo
• the pharmaceutically acceptable cations which can form a salt with one or both of the carboxyls (CO2M and CO2M-2) of the compounds of formula I are known to those skilled in the art. Examples include those where Ml and M2 independently can be alkali metals such as sodium, potassium and the like, ammonium and the like or hydrogen. It is noted that the compounds claimed in the instant invention are, as necessary, charged balanced in accordance with the knowledge of those skilled in the art.
• the pharmaceutically acceptable esterifying groups are such as would be readily apparent to a medicinal chemist, and include, for example, those described in detail in U.S. Pat. No. 4,309,438. Included within such pharmaceutically acceptable esters are those which are hydrolyzed under physiological conditions, such as pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl and methoxymethyl, and others described in detail in U.S. Pat. No. 4,479,947. These are also referred to as "biolabile esters”. Biolabile esters are biologically hydrolizable, and may be suitable for oral administration, due to good absorption through the stomach or intenstinal mucosa, resistance to gastric acid degradation and other factors.
• Some of the compounds of formula I may be crystallized or recrystallized from solvents such as organic solvents. In such cases solvates may be formed.
• This invention includes within its cope stoichiometric solvates including hydrates as well as compounds containing variable amounts of solvents such as water that may be produced by processess such as lyophilization.
• the compounds of formula I may be prepared in crystalline form by for example dissolution of the compound in water, preferably in the minimum quantity thereof, followed by admixing of this aqueous solution with a water miscible organic solvent such as a lower aliphatic ketone such as a di-(Ci-6) alkyl ketone, or a (Ci-6) alcohol, such as acetone or ethanol.
• R and/or R represents a to C 16 straight, branched or unsaturated alkyl group optionally substituted with 1 to 3 R x groups and optionally interrupted by one of the following O, S, SO2, -C(O)-, -C(O)-NRa-, -CO 2 - and all other variables are described as above.
• a subset of this invention is realized when R 1 and/or R is a C 5 to C 16 alkyl, preferably C 7 to C 16 alkyl and are not the same.
• a subset of compounds of formula I which is of interest relates to those compounds where M and M2 are independently hydrogen, sodium or potassium and all other variables are as described above.
• R 1 and/or R 2 represents C5-I6, preferably C7-I6 straight, branched or unsaturated alkyl optionally substituted with 1 to 2 R x , wherein all variables are as described above.
• R 1 and R 2 are wherein all other variables are described as above.
• Still another subset of compounds of formula I which is of interest relates to those compounds whe :rree RR 1 1 aanndd//oorr R 2 represents a group of the formula
• A is (CH2)l-5 and is phenyl, naphthyl, cyclohexyl or dibenzofuranyl.
• Still another subset of compounds of formula I that is of interest relates
• A is (CH2)l-3, A' is a bond, -O- or (CH2)l-2 and independently represent phenyl, thienyl, pyridyl, furanyl or cyclohexyl.
• Still another subset of compounds of formula I that is of interest relates to those compounds where one of R or R2 is:
• R x , M , and M are as originally defined.
• A is (CH2)l-2 and ⁇ is phenyl or cyclohexyl and the other of Rl or R2 is a group of the formula:
• A is (CH2)l-2, A is a single bond,
• ⁇ is phenyl or cyclohexyl and C ⁇ - C ⁇ - ⁇ ) is phenyl, thienyl or pyridyl.
• Still another subset of compounds of formula I that is of interest relates to those compounds where:
• R 1 is C5-7 alkyl substituted with 0 to 2 R x groups
• R2 is C7-I0 alkyl substituted with 0 to 2 R x groups
• R x A preferred subset of R x is RY.
• the compounds of the invention which are succinic acids or derivatives thereof, can be formulated in pharmaceutical compositions by combining the compound with a pharmaceutically acceptable carrier.
• a pharmaceutically acceptable carrier examples include metallo- ⁇ -lactamase inhibitory properties, and are useful when combined with a ⁇ -lactam antibiotic for the treatment of infections in animals, especially mammals, including humans.
• the compounds may be used, for example, in the treatment of infections of, amongst others, the respiratory tract, urinary tract and soft tissues and blood.
• the compounds may be employed in powder or crystalline form, in liquid solution, or in suspension. They may be administered by a variety of means; those of principal interest include: topically, orally and parenterally by injection (intravenously or intramuscularly).
• compositions for injection may be prepared in unit dosage form in ampules, or in multidose containers.
• the injectable compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain various formulating agents.
• the active ingredient may be in powder (lyophillized or non-lyophillized) form for reconstitution at the time of delivery with a suitable vehicle, such as sterile water.
• the carrier is typically comprised of sterile water, saline or another injectable liquid, e.g., peanut oil for intramuscular injections.
• various buffering agents, preservatives and the like can be included.
• Topical applications may be formulated in carriers such as hydrophobic or hydrophilic bases to form ointments, creams, lotions, in aqueous, oleaginous or alcoholic liquids to form paints or in dry diluents to form powders.
• Oral compositions may take such forms as tablets, capsules, oral suspensions and oral solutions.
• the oral composions may utilize carriers such as conventional formulating agents, and may include sustained release properties as well as rapid delivery forms.
• the compounds of the instant invention are metallo- ⁇ -lactamase inhibitors, which are intended for use in pharmaceutical compositions. Accordingly, it is preferable that the metallo- ⁇ -lactamase inhibitors are provided in substantially pure form, for example at least about 60% to about 75% pure, preferably about 85% to about 95% pure and most preferably about 98% or more pure (% are on a weight for weight basis). Impure preparations of the compounds may be used for preparing the more pure forms used in pharmaceutical compositions.
• the dosage to be administered depends to a large extent upon the condition and size of the subject being treated, the route and frequency of administration, the sensitivity of the pathogen to the particular compound selected, the virulence of the infection and other factors. Such matters, however, are left to the routine discretion of the physician according to principles of treatment well known in the antibacterial arts. Another factor influencing the precise dosage regimen, apart from the nature of the infection and peculiar identity of the individual being treated, is the molecular weight of the compound.
• compositions for human delivery per unit dosage may contain from about 0.01% to as high as about 99% of active material, the preferred range being from about 10-60%.
• the composition will generally contain from about 15 mg to about 2.5 g of the active ingredient; however, in general, it is preferable to employ dosage amounts in the range of from about 250 mg to 1000 mg.
• the unit dosage will typically include the pure compound in sterile water solution or in the form of a soluble powder intended for solution, which can be adjusted to neutral pH and isotonic.
• the invention described herein also includes a method of treating a bacterial infection in a mammal in need of such treatment comprising administering to said mammal a compound of formula I in conjunction with a ⁇ -lactam antibiotic such as a carbapenem, penicillin or cephalosporin in an effective combination.
• a ⁇ -lactam antibiotic such as a carbapenem, penicillin or cephalosporin in an effective combination.
• the preferred methods of administration of the Formula I compounds include oral and parenteral, e.g., i.v. infusion, i.v. bolus and i.m. injection.
• the compounds of formula I may suitably be administered to the patient at a daily dosage of from 0.7 to 50 mg/kg of body weight.
• a daily dosage of from 0.7 to 50 mg/kg of body weight For an adult human (of approximately 70 kg body weight), from 50 to 3000 mg, preferably from 100 to 1000 mg, of a compound according to the invention may be administered daily, suitably in from 1 to 6, preferably from 2 to 4, separate doses. Higher or lower dosages may, however, be used in accordance with clinical practice.
• the compounds may be used in combination with antibiotic agents for the treatment of infections caused by metallo- ⁇ -lactamase producing strains, in addition to those infections which are subsumed within the antibacterial spectrum of the antibiotic agent.
• Metallo- ⁇ -lactamase producing strains include: Bacillus cereus, Bacteroides fragilis, Aeromonas hydrophila, Klebsiella pneumoniae, Pseudomonas aeruginosa, Serratia marcescens, Stenotrophomonas maltoph ⁇ lia, Shigella flexneri, Legionella gormanii, Chryseobacterium meningosepticum, Chryseobacterium indologenes, Acinetobacter baumannii, Citrobacter freundii, and Aeromonas veronii.
• a compound of formula I in admixture or conjuction with a carbapenem, penicillin, cephalosporin or other ⁇ -lactam antibiotic or prodrug. It also advantageous to use a compound of formula I in combination with one or more ⁇ -lactam antibiotics because of the metallo- ⁇ -lactamase inhibitory properties of the compounds. In this case, the compound of formula I and the ⁇ -lactam antibiotic can be administered separately or in the form of a single composition containing both active ingredients.
• Carbapenems, penicillins, cephalosporins and other ⁇ -lactam antibiotics suitable for co-administration with the compounds of Formula I, whether by separate administration or by inclusion in the compositions according to the invention, include both those known to show instability to or to be otherwise susceptible to metallo- ⁇ -lactamases and also known to have a degree of resistance to metallo- ⁇ -lactamase.
• a dehydropeptidase (DHP) inhibitor may also be combined.
• DHP dehydropeptidase
• Many carbapenems are susceptible to attack by a renal enzyme known as DHP. This attack or degradation may reduce the efficacy of the carbapenem antibacterial agent.
• Inhibitors of DHP and their use with carbapenems are disclosed in, e.g., (European Patent 0007614, filed July 24, 1979 and application number 82107174.3, filed August 9, 1982.
• a preferred DHP inhibitor is 7-(L-2-amino-2-carboxyethylthio)-2-(2,2- dimethylcyclopropanecarboxamide)-2-heptenoic acid or a useful salt thereof.
• compounds of the present invention in combination with a carbapenem such as imipenem and a DHP inhibitor such as, cilastatin is contemplated within the scope of this invention.
• a serine ⁇ -lactamase inhibitor such as clavulanic acid, sulbactam or tazobactam may also be co-administered with the compound of the invention and ⁇ - lactam antibiotics, either by separate administration, or co-formulation with one, other or both of the compounds of the invention and the ⁇ -lactam antibiotic.
• carbapenems that may be co-administered with the compounds of formula I include imipenem, meropenem, biapenem, (4R, 5S, 6S)-3- [3 S , 5 S)-5-(3-carboxyphenyl-carbamoyl)pyrrolidin-3-ylthio] -6-( 1 R)- 1 -hydroxyethyl] - 4-methyl-7-oxo-l-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid, (IS, 5R, 6S)-2-(4- (2-(((carbamoylmethyl)-l ,4-diazoniabicyclo[2.2.2]oct- 1 -yl)-ethyl(l ,8- naphthosultam)methyl)-6-[ 1 (R)-hydroxyethyl] - 1 -methylcarbapen-2-em-3 -carboxylate chloride, BMS18
• penicillins suitable for co-administration with the compounds according to the invention include benzylpenicillin, phenoxymethylpenicillin, carbenicillin, azidocillin, propicillin, ampicillin, amoxycillin, epicillin, ticarcillin, cyclacillin, pirbenicillin, azloccillin, mezlocillin, sulbenicillin, piperacillin, and other known penicillins.
• the penicillins may be used in the form of pro-drugs thereof; for example as in vivo hydro lysable esters, for example the acetoxymethyl, pivaloyloxymethyl, ⁇ -ethoxycarbonyloxy-ethyl and phthalidyl esters of ampicillin, benzylpenicillin and amoxycillin; as aldehyde or ketone adducts of penicillins containing a 6- ⁇ -aminoacetamido side chain (for example hetacillin, metampicillin and analogous derivatives of amoxycillin); and as a- estsers of carbenicillin and ticarcillin, for example the phenyl and indanyl -esters.
• in vivo hydro lysable esters for example the acetoxymethyl, pivaloyloxymethyl, ⁇ -ethoxycarbonyloxy-ethyl and phthalidyl esters of ampicillin, benzylpenicillin and amoxy
• cephalosporins examples include, cefatrizine, cephaloridine, cephalothin, cefazolin, cephalexin, cephacetrile, cephapirin, cephamandole nafate, cephradine, 4- hydroxycephalexin, cephaloglycin, cefoperazone, cefsulodin, ceftazidime, cefuroxime, cefinetazole, cefotaxime, ceftriaxone, and other known cephalosporins, all of which may be used in the form of pro-drugs thereof.
• ⁇ -lactam antibiotics other than penicillins and cephalosporins that may be co-administered with the compounds according to the invention include aztreonam, latamoxef (Moxalactam-trade mark), and other known ⁇ -lactam antibiotics such as carbapenems like imipenem, meropenem or (4R, 5S, 6S)- 3-[(3S,5S)-5-(3-carboxyphenylcarbamoyl)pyrrolidin-3-ylthio]-6-(lR)-l- hydroxyethyl]-4-methyl-7-oxo-l-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid, all of which may be used in the form of pro-drugs thereof.
• aztreonam latamoxef (Moxalactam-trade mark)
• other known ⁇ -lactam antibiotics such as carbapenems like imipenem
• Preferred carbapenems are imipenem, meropenem and (4R, 5S, 6S)-3- [(3S,5S)-5-(3-carboxyphenylcarbamoyl)pyrrolidin-3-ylthio]-6-(lR)-l-hydroxyethyl]- 4-methyl-7-oxo-l-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid.
• penicillins for co-administration with the compounds according to the invention include ampicillin, amoxycillin, carbenicillin, piperacillin, azlocillin, mezlocillin, and ticarcillin.
• Such penicillins may be used in the form of their pharmaceutically acceptable salts, for example their sodium salts.
• ampicillin or amoxycillin may be used in the form of fine particles of the zwitterionic form (generally as ampicillin trihydrate or amoxycillin trihydrate) for use in an injectable or infusable suspension, for example, in the manner described herein in relation to the compounds of formula I.
• Amoxycillin for example in the form of its sodium salt or the trihydrate, is particularly preferred for use in compositions according to the invention.
• cephalosporins for co-administration with the compounds according to the invention include cefotaxime, ceftriaxone and ceftazidime, which may be used in the form of their pharmaceutically acceptable salts, for example their sodium salts.
• each unit dose may suitably comprise from about 25 to about 1000 mg, preferably about from 50 to about 500 mg, of a compound according to the invention.
• Each unit dose may, for example, be 62.5, 100, 125, 150, 200 or 250 mg of a compound according to the invention.
• the ratio of the amount of the compounds of formula I to the amount of the other ⁇ -lactam antibiotic may vary within a wide range. The said ratio may, for example, be from 100:1 to 1 : 100; more particularly, it may for example, be from 2:1 to 1 :30.
• the amount of carbapenem, penicillin, cephalosporin or other ⁇ -lactam antibiotic according to the invention will normally be approximately similar to the amount in which it is conventionally used.
• the claimed invention also includes the use of a compound of formula I, a pharmaceutically acceptable salt, ester, prodrug, anhydride or solvate thereof, in the manufacture of a medicament for the treatment of bacterial infections.
• the claimed invention also includes the use of a compound of formula I as a metallo- ⁇ -lactamase inhibitor.
• the claimed invention further includes a method of treating bacterial infections in humans or animals which comprises administering, in combination with a ⁇ -lactam antibiotic, a therapeutically effective amount of a metallo- ⁇ -lactamase inhibitor of formula I.
• the claimed invention further includes a method of treating bacterial infections in humans or animals which comprises administering, in combination with a carbapenem antibiotic, a therapeutically effective amount of a metallo- ⁇ -lactamase inhibitor of formula I.
• the claimed invention also includes a composition comprising a metallo- ⁇ -lactamase inhibitor of formula I together with a ⁇ -lactam antibiotic and a pharmaceutically acceptable carrier.
• the claimed invention also includes a composition comprising a metallo- ⁇ -lactamase inhibitor of formula I together with a carbapenem antibiotic and a pharmaceutically acceptable carrier.
• compositions discussed above may optionally include a serine ⁇ - lactamase inhibitor as described above as well as a DHP inhibitor.
• the compounds of the present invention are synthesized using the general conditions shown in the accompanying flow charts (A through E). FLOW SHEET A
• Ar 1 optionally substituted aryl or heteroaryl group
• Ar 2 optionally substituted aryl or heteroaryl
• P 1 carboxyl protecting group
• P 2 carboxyl protecting group
• the 2,3-disubstituted succinic acid compounds of the present invention can be prepared by the general methods described in Flow Sheets A-E.
• R x substituents When one or more R x substituents are present in a compound synthesized according to the Flow Sheets, it is sometimes advantageous that they be carried through the syntheses in protected or precursory form and then be deprotected or elaborated at or near the end of the synthesis.
• a desired R x group is incompatible with the reaction conditions of the synthesis being employed, said R x group may be introduced initially in a protected form and then be deprotected at the end of the synthesis.
• Flow Sheet A The synthesis of Flow Sheet A is based on a known literature procedure (M. J. Crimmin et. al., SynLett 1993, 137).
• the Rl -substituted acetic acid starting materials Al are readily available from commercial sources or are readily prepared by a variety of methods known in the art. Briefly, the starting material Al is alkylated with an ester derivative of bromoacetic acid, employing a chiral auxiliary group to achieve stereoselectivity in the reaction. After removal of the chiral auxiliary to give A4, the R2-alkyl group is introduced stereoselectively by an alkylation reaction to give A5. Removal of the carboxyl protecting group of A5 provides the final compound A6.
• the first step of Flow Sheet A is introduction of the chiral auxiliary.
• a suggested method is as follows.
• a mixed anhydride is formed between the starting carboxylic acid Al and pivalic acid by treating Al with a tertiary amine base such as triethylamine and pivaloyl chloride in a suitable ethereal solvent such as tetrahydrofuran at reduced temperature such as between -78°C and 0°C.
• intermediate A2 is obtained.
• Intermediate A2 is deprotonated with a strong base such as sodium hexamethyldisilazide in a solvent such as tetrahydrofuran at reduced temperature such as between -78°C and -70°C.
• the resulting enolate is alkylated by addition of BrCH2CO2Pl, where Pi is a removable carboxyl protecting group.
• compound A3 is obtained by conventional isolation and purification techniques.
• Suitable removable ester derivatives of bromoacetic acid for this alkylation reaction are t-butyl bromoacetate, allyl bromoacetate or benzyl bromoacetate.
• the oxazolidinone chiral auxiliary group of A3 is removed by a hydrolysis reaction.
• Aqueous lithium hydroxide and aqueous hydrogen peroxide are employed for this reaction along with an organic co-solvent such as tetrahydrofuran.
• the reaction is carried-out at a temperature of from 0°C to 30°C for a reaction time of from 30 min to 4 hours.
• acidification conventional isolation and purification provides intermediate A4.
• An alternative method of removing the chiral auxilliary consists of reacting A3 with lithium benzyloxide (LiOCEt ⁇ Ph) followed by cleavage of the resulting benzyl ester to give A4.
• reaction of A3 with lithium benzyloxide is carried-out in tetrahydrofuran as solvent at a temperature of from -78°C to 30°C for a reaction time of from 30 min to 4 hours.
• Cleavage of the resulting benzyl ester is accomplished in conventional fashion, eg by hydrogenolysis employing a suitable catalyst such as palladium on carbon in an appropriate solvent such as ethanol at 1-2 atmospheres pressure of hydrogen.
• compound A4 is obtained.
• Alkylation of A4 to give A5 is accomplished by deprotonating A4 with
• Preferred bases for this reaction are lithium bis(trimethylsilyl)amide and lithium diisopropylamide.
• the reaction is allowed to proceed at a temperature of from -78°C to 25°C for a reaction time of from 1 to 12 hours. Progress of the reaction can be monitored by conventional analytical methods, eg HPLC and TLC.
• Preferred alkylating agents for this reaction are alkyl iodides and alkyl bromides. Other suitable alkylating agents are well known in the art and include alkyl trifluoromethanesulfonates, alkyl methanesulfonates and alkyl tosylates.
• the minor stereoisomer produced in this reaction can often be separated from A5 at this stage by conventional chromatographic techniques. However, it is often preferable to carry-out this separation at the stage of A6, after removal of the carboxyl protecting group as described below. Removal of the carboxyl protecting group of A5 by standard methods gives the final compound A6.
• a strong acid such as trifluoroacetic acid in a suitable solvent such as dichloromethane.
• the reaction is carried-out at a temperature of from 0°C to 30°C for a reaction time of from 1 to 8 hours.
• the final compound A6 is then isolated by conventional techniques.
• a chiral auxiliary of the opposite absolute configuration eg. lithio-(4S)-benzyl-2- oxazolidinone
• Flow Sheet B illustrates a variation of Flow Sheet A which is preferred in certain cases, for example when Arl is a heteroaryl group such as pyridyl.
• the second substituent on the succinic acid is introduced by an aldol reaction instead of an alkylation reaction.
• the synthesis begins with compound A4, which is prepared as described in Flow Sheet A.
• Compound A4 is deprotonated with >2 equivalents of a strong hindered base to give a dianion which is then reacted with an aldehyde Arl CHO to give B15, where Arl 1S a n optionally substituted aryl or heteroaryl group, terms which are defined above.
• the deprotonation reaction is carried-out in a suitable solvent such as tetrahydrofuran at a temperature of from - 78°C to -70°C for a reaction time of from 30 min to 3 hours.
• Preferred bases for this reaction are lithium bis(trimethylsilyl)amide and lithium dusopropylamide. After addition of the aldehyde, the reaction is allowed to proceed at a temperature of from - 78°C to 25°C for a reaction time of from 1 to 12 hours. After conventional isolation and purification, intermediate Bl is obtained.
• Compound Bl is next cyclized to the lactone B2. Suitable conditions for this cyclization reaction would be exposure of Bl to acetic anhydride and triethylamine in an inert solvent such as dichloromethane. Reductive opening of lactone B2, such as by hydrogenolysis over palladium on carbon in a suitable solvent such as methanol, provides compound B3. Removal of the carboxyl protecting group of B3 by conventional methods then gives the final compound B4.
• Flow Sheet C illustrates an extension of the synthesis of Flow Sheet A which makes possible the introduction of a variety of preferred biaryl-type R2 substituents.
• the P2 carboxyl protecting group is introduced in conventional fashion.
• a preferred P2 group is p-methoxybenzyl which can be introduced employing p-methoxybenzyl alcohol, a carbodiimide reagent such as 1,3- diisopropylcarbodiimide and N,N-dimethylaminopyridine catalyst in a suitable inert solvent such as dichloromethane.
• suitable ester protecting groups known in the art could also be employed (see e.g. Greene, T. W., et al. Protective Groups in Organic Synthesis, John Wiley & Sons. Inc., 1991).
• the palladium catalyzed cross-coupling reaction between C2 and R3- Met is carried-out by procedures known in the scientific and patent literature.
• Met is a boronic acid moiety [-B(OH)2] the reaction is commonly known as a Suzuki reaction (see Suzuki, Chem. Rev. 1995, 95, 2457).
• Compound C2 is combined with the boronic acid R3-B(0H)2 in a coupling solvent such as 1,2-dimethoxyethane, N,N- dimethylformamide or toluene, optionally with water as a co-solvent, with a base such as sodium carbonate and a palladium catalyst such as tetrakis(triphenylphosphine)palladium(0).
• a coupling solvent such as 1,2-dimethoxyethane, N,N- dimethylformamide or toluene, optionally with water as a co-solvent
• a base such as sodium carbonate
• a palladium catalyst such as
• the reaction is carried-out at a temperature of from 20 °C to 125 °C for a reaction time of from 1 to 48 hours.
• the coupled product C3 is then isolated by conventional techniques.
• Met is a trialkyltin moiety
• the reaction is commonly known as a Stille reaction and the cross-coupling is carried-out by procedures well known in the literature (T. N. Mitchell, Synthesis 1992, 803).
• Flow Sheet D illustrates an alternative synthesis of compounds of the present invention.
• the synthesis of Flow Sheet D is based on known literature procedures (see for example J. L.
• a preferred strong base for the deprotonation reaction is lithium dusopropylamide.
• Suitable oxidizing agents for the synthesis of Flow Sheet D include iodine, copper(II) salts such as CuBr2, and titanium tetrachloride.
• Flow Sheet D Since the synthesis of Flow Sheet D is based on a dimerization-type reaction, it is best suited for the synthesis of symmetrically 2,3-disubstituted succinic acids. For this reason, it is generally less preferred than the syntheses of Flow Sheets A, B and C.
• the synthesis of Flow Sheet D also generally produces a racemic mixture of stereoisomers.
• reaction mixture was partitioned between ethyl acetate and sat. aqueoues NH4CI and the organic phase was washed with water and brine. The organic layer was dried over Na2SO4 and evaporated in vacuo to give a semi-solid. This crude material was triturated with 10 ml CH2CI2 and filtered through a sintered-glass funnel.
• EVIP-1 metallo- ⁇ -lactamase lacking the N-terminal 18 hydrophobic amino acids which encode the putative periplasmic signal sequence (EMBL access code PACATAAC6) was PCR amplified from plasmid DNA prepared from a carbapenem-resistant strain of Pseudomonas aeruginosa (CL5673). The PCR product was cloned into pET30a+ (Novegen) and expressed in E.coli BL21(DE3) after induction with 0.5 mM IPTG for 20 hours at room temperature in minimal media supplemented with casamino acids and 348 ⁇ M ZnSO 4 . Soluble IMP-1 was purified from cell extracts by SP-Sepharose (Pharmacia) ion exchange and Superdex 75 (Pharmacia) size-exclusion chromatography.
• the IC 50 of succinate derivatives of Formula I was determined following a 15 minute incubation at 37°C with EVIP-1 (0.75nM in 50mM MOPS, pH 7). Using initial velocity as a measure of activity, inhibition was monitored spectrophotometrically at 490nm in a Molecular Devices SPECTRAmaxTM 250 96- well plate reader employing nitrocefin as the reporter substrate at approximately K m concentration (60 ⁇ M). A laboratory strain of E.coli engineered to express EVIP-1 was used to evaluate the ability of succinate derivatives of Formula I to reverse metallo- ⁇ - lactamase-mediated carbapenem resistance in bacteria.
• Native EVIP-1 which included the N-terminal periplasmic signal sequence, was PCR amplified from CNA isolated from a carbapenem resistant P. aeruginosa clinical isolate, CL56673, and cloned into the pET30a vector. The basal (uninduced) level of EVIP-1 expressed when pET30a- EVIP-1 was introduced into E.
• coli BL21(DE3) resulted in 4-, 64- or 500-fold reduced sensitivity to impenem, meropenem or (lS,5R,6S)-l-methyl-2- ⁇ 7-[4- (aminocarbonylmethyl)-l,4-diazoniabicyclo(2.2.2)octan-l-yl]methyl-fluoren-9-on-3- yl ⁇ -6-(lR-hydroxyethyl)-carbapen-2-em-3-carboxylate chloride (a carbapenem synthesized at Merck Research Laboratories) respectively.
• the minimum inhibitory concentration (MIC) of (lS,5R,6S)-l-methyl-2- ⁇ 7-[4- (aminocarbonylmethyl)-l,4-diazoniabicyclo(2.2.2)octan-l-yl]methyl-fluoren-9-on-3- yl ⁇ -6-(lR-hydroxyethyl)-carbapen-2-em-3-carboxylate chloride was typically increased from 0.06-0.12 ⁇ g/ml to 16-32 ⁇ g/ml by the expression of EVIP-1.
• E an overnight culture of E.
• coli BL2(DE3)/pET30a-EVIP-l grown 35°C in LB broth (Difco) or Mueller Hinton broth (BBL) supplemented with kanamycin (50 ⁇ M/ml), was diluted to a final concentration of ⁇ 10 5 cells/ml in Mueller Hinton broth (BBL) containing a subinhibitory concentration (0.25x MIC) of the carbapenem, (lS,5R,6S)-l-methyl-2- ⁇ 7-[4-(aminocarbonylmethyl)-l,4- diazoniabicyclo(2.2.2)octan-l-yl]methyl-fluoren-9-on-3-yl ⁇ -6-(lR-hydroxyethyl)- carbapen-2-em-3-carboxylate chloride.
• Representative compounds of Formula I were tested as inhibitors against purified EVIP-1 metallo- ⁇ -lactamase and found to be active in an IC 50 range of from about 0.2nM to about 500 ⁇ M. The ability of representative compounds of
• Compounds of Formula I in the concentration range of from about 0.002 ⁇ M to about lOO ⁇ M.

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